دورية أكاديمية
Protease-activated receptor-2 (PAR-2): structure-function study of receptor activation by diverse peptides related to tethered-ligand epitopes.
| العنوان: | Protease-activated receptor-2 (PAR-2): structure-function study of receptor activation by diverse peptides related to tethered-ligand epitopes. |
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| المؤلفون: | Maryanoff BE; Drug Discovery, The R.W. Johnson Pharmaceutical Research Institute, Spring House, Pennsylvania 19477, USA. bmaryano@prius.jnj.com, Santulli RJ, McComsey DF, Hoekstra WJ, Hoey K, Smith CE, Addo M, Darrow AL, Andrade-Gordon P |
| المصدر: | Archives of biochemistry and biophysics [Arch Biochem Biophys] 2001 Feb 15; Vol. 386 (2), pp. 195-204. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: United States NLM ID: 0372430 Publication Model: Print Cited Medium: Print ISSN: 0003-9861 (Print) Linking ISSN: 00039861 NLM ISO Abbreviation: Arch Biochem Biophys Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: <2000- > : San Diego, CA : Elsevier Original Publication: New York, NY : Academic Press |
| مواضيع طبية MeSH: | Peptides/*chemistry , Peptides/*pharmacology , Receptors, Thrombin/*agonists , Receptors, Thrombin/*metabolism, Amino Acid Motifs ; Amino Acid Sequence ; Amino Acid Substitution ; Animals ; Blood Platelets/drug effects ; Blood Platelets/metabolism ; Blood Platelets/physiology ; Calcium/metabolism ; Calcium Signaling/drug effects ; Cell Line ; Directed Molecular Evolution ; Dose-Response Relationship, Drug ; Drug Design ; Humans ; Ligands ; Mice ; Peptide Library ; Peptides/genetics ; Peptides/metabolism ; Platelet Aggregation/drug effects ; Rats ; Receptor, PAR-1 ; Receptor, PAR-2 ; Receptors, Thrombin/chemistry ; Receptors, Thrombin/genetics ; Structure-Activity Relationship |
| مستخلص: | Protease-activated receptor-2 (PAR-2) is a tethered-ligand, G-protein-coupled receptor that is activated by proteolytic cleavage or by small peptides derived from its cleaved N-terminal sequence, such as SLIGRL-NH2. To assess specific PAR activity, we developed an immortalized murine PAR-1 (-/-) cell line transfected with either human PAR-2 or PAR-1. A "directed" library of more than 100 PAR agonist peptide analogues was synthesized and evaluated for PAR-2 and PAR-1 activity to establish an in-depth structure-function profile for specific action on PAR-2. The most potent agonist peptides (EC50 = 2-4 microM) had Lys at position 6, Ala at position 4, and pFPhe at position 2; however, these also exhibited potent PAR-1 activity (EC50 = 0.05-0.35 microM). We identified SLIARK-NH2 and SL-Cha-ARL-NH2 as relatively potent, highly selective PAR-2 agonists with EC50 values of 4 microM. Position 1 did not tolerate basic, acidic, or large hydrophobic amino acids. N-Terminal capping by acetyl eliminated PAR-2 activity, although removal of the amino group reduced potency by just 4-fold. At position 2, substitution of Leu by Cha or Phe gave equivalent PAR-2 potency, but this modification also activated PAR-1, whereas Ala, Asp, Lys, or Gln abolished PAR-2 activity; at position 3, Ile and Cha were optimal, although various amino acids were tolerated; at position 4, Ala or Cha increased PAR-2 potency 2-fold, although Cha introduced PAR-1 activity; at position 5, Arg or Lys could be replaced successfully by large hydrophobic amino acids. These results with hexapeptide C-terminal amides that mimic the native PAR-2 ligand indicate structural modes for obtaining optimal PAR-2 activity, which could be useful for the design of PAR-2 antagonists. |
| المشرفين على المادة: | 0 (Ligands) 0 (Peptide Library) 0 (Peptides) 0 (Receptor, PAR-1) 0 (Receptor, PAR-2) 0 (Receptors, Thrombin) SY7Q814VUP (Calcium) |
| تواريخ الأحداث: | Date Created: 20010523 Date Completed: 20010607 Latest Revision: 20131121 |
| رمز التحديث: | 20240627 |
| DOI: | 10.1006/abbi.2000.2207 |
| PMID: | 11368342 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 0003-9861 |
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| DOI: | 10.1006/abbi.2000.2207 |